Fig. 3

EYA4 inhibited proliferation, clonogenicity and invasiveness of HCC cells by the suppression of RAP1. a Western blotting analyses showing the RAP1 protein expression levels in pEYA4 transfectants (left: Huh-7, right: PLC5) after transfection with or without Flag-tagged human RAP1A expressing plasmid (Flag-RAP1A). b–d Effects of RAP1 expression on cell proliferation (b), clonogenicity (c) and cell invasiveness (d) of EYA4-overexpressing HCC cells. pEYA4 transfectants were transfected with or without Flag-RAP1A, and the resulting cell proliferation, clonogenic outgrowth and cell invasiveness were determined by CCK-8 assays, colony formation assays and transwell invasion assays, respectively. *P < 0.05 between groups. e Western blotting analyses of the RAP1 expression levels in Huh-7 (left panel) and PLC5 (right panel) with or without Flag-RAP1A transfection. f–h Effects of RAP1 expression on cell proliferation (f), clonogenicity (g) and cell invasiveness (h) of HCC cells. The effects were analyzed by CCK-8 assays, colony formation assays and transwell invasion assays, respectively. *P < 0.05 versus the Flag-RAP1A group. EYA4 eyes absent homolog 4, Flag-RAP1A Flag-tagged human RAP1A expressing plasmid, Flag-Vector Flag-tagged vector plasmid, GAPDH glyceraldehyde 3-phosphate dehydrogenase, pEYA4 EYA4-expressing plasmid, RAP1 RAS-related protein 1, RAP1A RAS-related protein 1A gene, Un untransfected control